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cAMP responsive element binding protein-1 is a transcription factor of lysosomal-associated protein transmembrane-4 Beta in human breast cancer cells.

Zhang M, Xu JJ, Zhou RL, Zhang QY - PLoS ONE (2013)

Bottom Line: Lysosomal-associated protein transmembrane-4 beta (LAPTM4B) is a potential proto-oncogene, whose overexpression is involved in cancer occurrence and progression.However, its transcriptional regulation mechanism is still unclear.The +10+292 promoter region was possessed the highest transcriptional activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical laboratory, Key laboratory of Carcinogenesis and Translational Research (Ministry of Education), Peking University Cancer Hospital & Institute, Beijing, China.

ABSTRACT
Lysosomal-associated protein transmembrane-4 beta (LAPTM4B) is a potential proto-oncogene, whose overexpression is involved in cancer occurrence and progression. Its transcript is up-regulated in various types of solid tumors including breast cancer. However, its transcriptional regulation mechanism is still unclear. To investigate the mechanism of transcriptional regulation of LAPTM4B in human breast cancer cells, a series of luciferase reporter constructs and construct with mutated binding site for cAMP responsive element binding protein-1 (CREB1) were generated by PCR amplification and transiently transfected into breast cancer cells to determine the transcriptional activities of different promoter regions. The +10+292 promoter region was possessed the highest transcriptional activity. The ability of CREB1 to bind the LAPMT4B promoter was confirmed by electrophoretic mobility shift assay, super-shift and RNA interference experiments. Our study identified the core promoter region responsible for constitutive expression of LAPTM4B and clarified that CREB1 played an important role in LAPTM4B transcriptional regulation in human breast cancer cells.

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Related in: MedlinePlus

The upper panel is a mode chart of the 5′-genomic structure of LAPTM4B gene.Exon1 are depicted as a closed box, and 5′-region is black lines. The transcription start site depicted as right-point arrow. the gray box indicates the CREB1 binding site. The curly brace depicts the initiation codon in exon1. The lower panel shows the luciferase constructs ranging across the 5′-region. Closed boxes depict the luciferase. The CREB1 conserved binding site depicted as the gray box. Asterisk was referred to the mutated CREB1 binding site. (*P<0.05, n = 3).
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pone-0057520-g002: The upper panel is a mode chart of the 5′-genomic structure of LAPTM4B gene.Exon1 are depicted as a closed box, and 5′-region is black lines. The transcription start site depicted as right-point arrow. the gray box indicates the CREB1 binding site. The curly brace depicts the initiation codon in exon1. The lower panel shows the luciferase constructs ranging across the 5′-region. Closed boxes depict the luciferase. The CREB1 conserved binding site depicted as the gray box. Asterisk was referred to the mutated CREB1 binding site. (*P<0.05, n = 3).

Mentions: In order to determine the authenticity of the plasmids, all the constructs were digested by restriction endonucleases Hind III and Xho I. The digested products were electrophoresis in 2% gel agarose. Fig. 2 is a mode chart of the 5′-genomic structure of LAPTM4B gene and luciferase constructs.


cAMP responsive element binding protein-1 is a transcription factor of lysosomal-associated protein transmembrane-4 Beta in human breast cancer cells.

Zhang M, Xu JJ, Zhou RL, Zhang QY - PLoS ONE (2013)

The upper panel is a mode chart of the 5′-genomic structure of LAPTM4B gene.Exon1 are depicted as a closed box, and 5′-region is black lines. The transcription start site depicted as right-point arrow. the gray box indicates the CREB1 binding site. The curly brace depicts the initiation codon in exon1. The lower panel shows the luciferase constructs ranging across the 5′-region. Closed boxes depict the luciferase. The CREB1 conserved binding site depicted as the gray box. Asterisk was referred to the mutated CREB1 binding site. (*P<0.05, n = 3).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585361&req=5

pone-0057520-g002: The upper panel is a mode chart of the 5′-genomic structure of LAPTM4B gene.Exon1 are depicted as a closed box, and 5′-region is black lines. The transcription start site depicted as right-point arrow. the gray box indicates the CREB1 binding site. The curly brace depicts the initiation codon in exon1. The lower panel shows the luciferase constructs ranging across the 5′-region. Closed boxes depict the luciferase. The CREB1 conserved binding site depicted as the gray box. Asterisk was referred to the mutated CREB1 binding site. (*P<0.05, n = 3).
Mentions: In order to determine the authenticity of the plasmids, all the constructs were digested by restriction endonucleases Hind III and Xho I. The digested products were electrophoresis in 2% gel agarose. Fig. 2 is a mode chart of the 5′-genomic structure of LAPTM4B gene and luciferase constructs.

Bottom Line: Lysosomal-associated protein transmembrane-4 beta (LAPTM4B) is a potential proto-oncogene, whose overexpression is involved in cancer occurrence and progression.However, its transcriptional regulation mechanism is still unclear.The +10+292 promoter region was possessed the highest transcriptional activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical laboratory, Key laboratory of Carcinogenesis and Translational Research (Ministry of Education), Peking University Cancer Hospital & Institute, Beijing, China.

ABSTRACT
Lysosomal-associated protein transmembrane-4 beta (LAPTM4B) is a potential proto-oncogene, whose overexpression is involved in cancer occurrence and progression. Its transcript is up-regulated in various types of solid tumors including breast cancer. However, its transcriptional regulation mechanism is still unclear. To investigate the mechanism of transcriptional regulation of LAPTM4B in human breast cancer cells, a series of luciferase reporter constructs and construct with mutated binding site for cAMP responsive element binding protein-1 (CREB1) were generated by PCR amplification and transiently transfected into breast cancer cells to determine the transcriptional activities of different promoter regions. The +10+292 promoter region was possessed the highest transcriptional activity. The ability of CREB1 to bind the LAPMT4B promoter was confirmed by electrophoretic mobility shift assay, super-shift and RNA interference experiments. Our study identified the core promoter region responsible for constitutive expression of LAPTM4B and clarified that CREB1 played an important role in LAPTM4B transcriptional regulation in human breast cancer cells.

Show MeSH
Related in: MedlinePlus